P53对涎腺腺样囊性癌细胞端粒酶活性的抑制作用  

Inhibition of telomerase activity in salivary adenoid cystic carcinoma cells by P53

作　　者：闫炳智[1] 王洁[1] 张波[2] 董福生[1] 侯琳[2] 王旭[1] 

机构地区：[1]河北医科大学口腔医学院病理室,河北石家庄050017 [2]北京大学医学部病理系,北京100083

出　　处：《基础医学与临床》2005年第3期204-208,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30271422);河北省自然科学基金(C2004000624);国家留学人员科技活动择优资助项目

摘　　要：目的为了探讨P53基因对腺样囊性癌细胞端粒酶活性的抑制作用及机制。方法构建携带人野生型P53基因的腺病毒表达载体,以脂质体法瞬时转染腺样囊性癌SACC83细胞,RT PCR检测转染细胞P53基因mRNA的表达,TRAP PCR ELISA法检测转染细胞端粒酶活性的改变。并将P53基因与含有hTERT启动子核心调控区的荧光素酶报告基因pGL2630共转染SACC83细胞,测定转染细胞荧光素酶报告基因活性。结果1.瞬时转染含人野生型P53基因的重组腺病毒表达载体pΔE1P53于腺样囊性癌SACC83细胞后,其P53基因mRNA的表达明显增强;2.基因转染后,SACC83细胞内源性端粒酶活性显著降低。3.P53基因与含有hTERT启动子核心调控区的荧光素酶报告基因pGL2630共转染SACC83细胞后,其荧光素酶活性显著下降。结论外源性表达P53基因可以降低腺样囊性癌细胞SACC83细胞端粒酶活性,并可能通过抑制端粒酶hTERT基因启动子的转录活性实现。Objective To investigate the effect of P53 in telomerase activity in salivary adenoid cystic carcinoma cells. Human wild-type P53 gene was constructed into pΔE1 vector. Methods Liposome-mediated gene transfection method was used to transfer P53 gene into SACC-83 cells. The expression of P53 mRNA was assessed by reverse transcription polymerase chain reaction(RT-PCR) and the activity of telomerase was examined by TRAP-PCR-ELISA. pΔE1-P53 and hTERT promoter-luciferase reporter vector pGL2-630 were co-transfected into SACC-83 cells, and then the luciferase activity was examined. Results 1. The level of P53 mRNA increased after pΔE1-P53 transfection; 2. The activity of telomerase decreased obviously. 3. The luciferase activity was also depressed after the co-transfection of pΔE1-P53 and pGL2-630. Conclusion The results suggested that P53 gene may depress the activity of telomerase in salivary adenoid cystic carcinoma cell through inhibiting the transcriptional activity of hTERT promoter.

关 键 词：细胞端粒酶活性 抑制作用 涎腺腺样囊性癌 SACC-83 野生型P53基因 荧光素酶报告基因 hTERT启动子 腺病毒表达载体 RT-PCR检测 ELISA法检测 端粒酶HTERT 基因mRNA 腺样囊性癌细胞 荧光素酶活性 瞬时转染 转染细胞 基因启动子 脂质体法 基因活性 基因转染 转录活 

分 类 号：R739.8[医药卫生—肿瘤]